Method and composition of thyroid hormone analogues and nanoformulations thereof for treating anti-inflammatory disorders

ABSTRACT

Thyroid hormone antagonists and their nanoparticle formulations (Nanotetrac™ or Nanotriac™) act at a cell surface receptor to block angiogenesis and tumor cell proliferation. The complex anti-angiogenic performs actions on specific cytokines and chemokines. Thyroid hormone antagonists inhibit expression in tumor cells of cytokine genes, e.g., specific interleukins, and chemokine genes, such as fractalkine (CX3CL1), and chemokine receptor genes (CX3CR1) that are targets in the development of inflammation-suppressant drugs. This application discloses a novel composition of Tetra or Tri-iodothyroacetic acid (tetrac or triac), other thyroid partial agonists or antagonists and their nanoparticle formulations conjugated to polymers and encapsulating non-steroidal anti-inflammatory, anti-inflammatory glucocorticoids, and/or polyphenols for the management of various acute and chronic inflammatory disorders ranging from neurological, vascular, and musculoskeletal disorders.

RELATED APPLICATIONS

This application claims the benefit of U.S. Application No. 61/807,123,filed Apr. 1, 2013, the content of which is incorporated herein byreference.

FIELD OF TECHNOLOGY

The following relates to polymer conjugate forms of thyroid hormoneanalogues and derivatives thereof. More specifically the followingdisclosure relates to embodiments of nanoparticulate tri-iodothyroaceticor tetra-iodothyroacetic acid, alone or in combination with a secondagent, such as non-steroidal anti-inflammatory drugs, salicylates,anti-inflammatory glucocorticoids, polyphenols, for management ofinflammatory conditions. The disclosure also relates generally tomethods of using said embodiments including embodiments to decrease orcontrol inflammation responses by the subject.

BACKGROUND

Thyroid hormones, such as L-thyroxine (T4) and 3, 5,3′-triiodo-L-thyronine (T3), and their analogues such as GC-1, DITPA,tetraiodothyroacetic acid (tetrac) and triiodothyroacetic acid (triac),regulate many different physiological processes in different tissues invertebrates. It was previously known that many of the actions of thyroidhormones are mediated by the thyroid hormone receptor (“TR”) and a novelcell surface receptor for thyroid hormone (L-thyroxine, T4; T3) integrinαvβ3, at or near the Arg-Gly-Asp (RGD) recognition site on the integrin.The integrin receptor is not a homologue of the nuclear thyroid hormonereceptor (TR), but activation of this cell surface receptor results in anumber of nucleus-mediated events. A detailed description of the thyroidhormones, analogs thereof and their properties have been fully discussedand disclosed in US Patent Publication No. 2011/0052715A1 to Davis etal., U.S. Pat. No. 7,785,632 to Mousa et al., and U.S. Pat. No.8,668,926 to Mousa et al., incorporated by reference in their entiretyherein.

Evidence that thyroid hormone can act primarily outside the cell nucleushas come from studies of mitochondrial responses to T3 anddiiodothyronine (T2), from rapid onset effects of the hormone at thecell membrane, and from actions on cytoplasmic proteins. The recentdescription of a plasma membrane receptor for thyroid hormone onintegrin αvβ3 has provided some insight into effects of the hormone onmembrane ion pumps, such as the Na+/H+ anti porter, and has led to thedescription of interfaces between actions initiated at the membranethyroid hormone receptor and nuclear events that underlie importantcellular or tissue processes, such as, for example, angiogenesis andproliferation of certain tumor cells.

Inflammation is closely linked to cancer. Chronic inflammation increasesthe risk for various cancers, indicating that eliminating inflammationmay represent a valid strategy for cancer prevention and therapy. Thereis data suggesting that inflammation plays a role in the establishment,progression, and/or aggressiveness of various malignancies. As a tumordevelops, it expresses phenotypes similar to inflammatory cells.Molecular mediators and their respective receptors have a significantimpact on angiogenesis, cell migration, and metastasis. Given its myriadpro-tumor effects, inflammation has become a target for cancerprevention and therapy. COX-2 (cyclooxygenase 2, PTGS2) is the mostfrequently evaluated anti-cancer anti-inflammatory target, althoughnumerous other targets, such as NF-kB, cytokines/cytokine receptors,chemokines/chemokine receptors, FGF/FGFR (fibroblast growthfactor/receptor), and VEGF have also been examined. While initialstudies focused on various broad-spectrum NSAIDs (which non-specificallyinhibit both COX-1 and COX-2), more recent studies have examined COX-2specific agents, such as celecoxib. However, given the GI toxicity andnon-specific activity of NSAIDs, and the cardio-toxicity of specificCOX-2 inhibitors, the use of such agents remains controversial.Therefore, a need exists for the combined use of an effectiveanti-cancer agent, anti-angiogenic agent and an anti-inflammatory agentcapable of being selectively targeted to the tumor cells and reducesinflammation while reducing the toxicity caused by unselectiveanti-inflammatory agents.

SUMMARY

A first embodiment of this disclosure relates generally to a compositioncomprising a thyroid hormone antagonist conjugated to a polymer and atleast one anti-inflammatory agent encapsulated within the polymer,wherein said at least one anti-inflammatory agent is selected from anon-steroidal anti-inflammatory drug (NSAID), a salicylate, ananti-inflammatory glucocorticoid, and pirfenidone.

A second embodiment of this disclosure relates generally to a method fortreating an inflammatory or musculoskeletal condition comprising thesteps of conjugating thyroid hormone analogue to a polymer, forming aconjugated thyroid hormone analog, encapsulating inside the polymer ofthe conjugated thyroid hormone analogue at least one of a non-steroidalanti-inflammatory drug (NSAID), a salicylate, an anti-inflammatoryglucocorticoid, and pirfenidone and binding the conjugated thyroidhormone analogue to at least one chemokine receptor, cytokine receptor,interleukin or a combination of receptors thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a graphical representation of the effects ofnanoparticulate tetrac treatment on the differentially regulatedinterleukin genes of human breast cancer MDA-MB-231 cells.

FIG. 2 depicts a graphical representation of the effects ofnanoparticulate tetrac on the expression of a plurality of chemokineligands and receptors in MDA-MBA-231 cells.

FIG. 3 depicts a representation of the synthesis of thyroid hormoneconjugated to a polymer encapsulating an NSAID and/or polyphenol.

FIG. 4 depicts a graphical representation of the size distribution oftetrac conjugated PLGA nanoparticles encapsulating a polyphenol,resveratrol.

DETAILED DESCRIPTION OF THE INVENTION

A detailed description of the hereinafter described embodiments of thedisclosed apparatus and method are presented herein by way ofexemplification and not limitation with reference to the Figures.Although certain embodiments are shown and described in detail, itshould be understood that various changes and modifications may be madewithout departing from the scope of the appended claims. The scope ofthe present disclosure will in no way be limited to the number ofconstituting components, the materials thereof, the shapes thereof, therelative arrangement thereof, etc., and are disclosed simply as anexample of embodiments of the present disclosure.

As a preface to the detailed description, it should be noted that, asused in this specification and the appended claims, the singular forms“a”, “an” and “the” include plural referents, unless the context clearlydictates otherwise.

Embodiments of the novel compound of this present disclosure may beformulated using a thyroid hormone analogue as a targeting agent and/orinflammation suppressing agent. The targeting agent may direct the novelcompound toward the tumor site and/or the site of inflammation. Examplesof thyroid hormone analogues are also provided herein and can includetriiodothyronine (T3), levothyroxine (T4), T4 or T3 N-Methyl, T4 or T3N-Ethyl, T4 or T3 N-Triphenyl, T4 or T3 N-Propyl, T4 or T3 N-Isopropyl,T4 or T3 N-tertiary butyl,3,5-dimethyl-4-(4′-hydroy-3′-isopropylbenzyl)-phenoxy acetic acid(GC-1), or 3,5-diiodothyropropionic acid (DITPA), tetraiodothyroaceticacid (TETRAC), and triiodothyroacetic acid (TRIAC), additionalantagonists described below and in Table 1 and pharmaceuticallyacceptable salts thereof.

“Pharmaceutically acceptable salts” may refer to pharmaceuticallyacceptable salts of thyroid hormone analogues, polymeric forms, andderivatives, which salts are derived from a variety of organic andinorganic counter ions well known in the art and include, by way ofexample only, sodium, potassium, calcium, magnesium, ammonium,tetra-alkyl ammonium, and the like; and when the molecule contains abasic functionality, salts of organic or inorganic acids, such ashydrochloride, hydro-bromide, tartrate, mesylate, acetate, maleate,oxalate and the like can be used as the pharmaceutically acceptablesalt. The term also includes both acid and base addition salts.

The compounds described herein, or their pharmaceutically acceptablesalts, may have asymmetric carbon atoms in their structure. Thecompounds disclosed herein and their pharmaceutically acceptable saltsmay therefore exist as single enantiomers, diastereoisomers, racemates,and mixtures of enantiomers and diastereomers. All such singleenantiomers, diastereoisomers, racemates and mixtures thereof areintended to be within the scope of this invention. Absoluteconfiguration of certain carbon atoms within the compounds, if known,may be indicated by the appropriate absolute descriptor R or S.

In some embodiments, the thyroid hormone analogue may be ananti-angiogenic thyroid hormone analogue, also referred to as a thyroidhormone antagonist. A thyroid hormone analogue may include substancesthat block L-T3 or L-T4 at the integrin alpha v beta 3 receptors (αvβ3).The terms “anti-angiogenesis” or “anti-angiogenic” may refer to anycompound or substance that inhibits or antagonizes angiogenesis, whetheralone or in combination with another substance. Examples of thyroidhormone antagonists may include, but are not limited to,tetraiodothyroacetic acid (tetrac), triiodothyroacetic acid (triac),phthalates, desethylamiodarone, NH-3, sulfonyl nitrophenyl thiazides,DHPPA, and the additional examples shown in Table 1 below.

TABLE 1 THYROID ANTAGONISTS EXAMPLES A.

Tetrac B.

DIBRT C.

NH-3 D.

E.

1-850 F.

G.

H.

In some embodiments, the thyroid hormone analogue may be conjugated to apolymer. The conjugation between the polymer and the thyroid hormoneanalogue may occur via a covalent or non-covalent bond, depending on thepolymer being used. In some embodiments, the polymer conjugation mayoccur through an ester linkage, anhydride linkage, ether linkage orsulfhydryl linkage immobilizing the thyroid hormone analogue to thesurface of the polymer. In some embodiments, the linkage may include alinker between 3 and 15 atoms long. In alternative embodiments, thelinker may be between 3-4, 3-5, 3-6, 3-7 or 3-8 atoms long. The linkerbetween the thyroid hormone analogue and the polymer may be attached onthe outer ring hydroxyl group of the thyroid hormone analog. The thyroidhormone analogue conjugated to a polymer described above may be alsoreferred to as a “conjugated thyroid hormone analog.”

EXAMPLE 1 Thyroid Hormone Conjugated to a Polymer Via an Ester Linkage

The polymer conjugations may be used to improve drug viability. Whilemany old and new therapeutics are well-tolerated, many compounds mayneed advanced drug technologies to decrease toxicity, increasecirculatory time, or modify biodistribution. One strategy for improvingdrug viability is the utilization of water-soluble polymers. Variouswater-soluble polymers have been shown to modify biodistribution,improve the mode of cellular uptake, change the permeability throughphysiological barriers, and modify the rate of clearance through thebody. To achieve either a targeting or sustained-release effect,water-soluble polymers have been synthesized that contain drug moietiesas terminal groups, as part of the backbone, or as pendent groups on thepolymer chain as well as encapsulating additional compounds inside thepolymer to control distribution thereof.

In some embodiments, the polymer that may be conjugated to the thyroidhormone analogue may include but is not limited to polylactic acid(PLLA), polyglycolic acid (PGA), poly(lactic-co-glycolic acid) (PLGA),polyacrylic acid, polyethylene glycol (PEG), poly-L-lysine, chitosan,hyaluronic acid, fatty acids, polyamine, polyvinyl alcohol, acrylic acidethylene co-polymer, methoxy polyethylene, human serum albumin,polyethylene oxide (PEO), cellulose derivatives, folate linkedcyclodextrin, folate linked cyclodextran, alginate, carrageenan, pectin,dextran, collagen, polyaniline, polyalanine, polytryptophan,polytyrosine, co-polymers and combinations thereof.

For example, in one embodiment, a polymer conjugation may be formedthrough an ester linkage using polyvinyl alcohol. In this preparationcommercially available polyvinyl alcohol (or related co-polymers) may beesterified by treatment with the acid chloride of thyroid hormoneanalogues, including the acid chloride form. The hydrochloride salt maybe neutralized by the addition of triethylamine to form triethylaminehydrochloride which can be washed away with water upon precipitation ofthe thyroid hormone ester polymer form for different analogues.

In an alternative example, a polymer conjugation through an anhydridelinkage using acrylic acid ethylene co-polymer similar to the previouspolymer covalent conjugation may be used, however, an anhydride linkagethat is derived from reaction of an acrylic acid co-polymer may beformed. This anhydride linkage may be susceptible to hydrolysis in vivoto release the thyroid hormone analog. Neutralization of thehydrochloric acid may be accomplished by treatment with triethylamineand subsequent washing of the precipitated polyanhydride polymer withwater which removes the triethylamine hydrochloride byproduct. Thisreaction may lead to the formation of thyroid hormone analogue acrylicacid co-polymer+triethylamine. Upon in vivo hydrolysis, the thyroidhormone analogue may be released over a time frame that can becontrolled and manipulated.

In an alternative embodiment, the polymer may be conjugated to thecarboxylic acid or the hydroxyl group of the thyroid hormone analogue asdepicted in example 2 and example 3 below.

EXAMPLE 2 Route of Tetrac/Polymer Conjugates Synthesis Via CarboxylicAcid Group

EXAMPLE 3 Route of Tetrac/Polymer Conjugates Synthesis Via HydroxylGroup

In alternative embodiments, a variety of synthetic, natural andbiopolymeric side groups with efficient biodegradable backbone polymersmay be conjugated to the thyroid hormone analogues. These alternativepolymers may include Poly alkyl glycols, polyesters, poly anhydride,poly saccharide, and poly amino acids. Example may include bi-functionalPEG, methoxy-PEG, polylactic-co-lysine and polyamidoamine.

Furthermore, in some embodiments, the polymer may be formulated into amicroparticle or nanoparticle. As used herein, the term “nanoparticle”refers to particles between about 1 nm and less than 1000 nm indiameter. In suitable embodiments, the diameter of the nanoparticles ofthe present invention may have a particle size having a diameter betweenapproximately 10 nm to <1000 nm. In other embodiments, the particle maybe less than 500 nm in diameter, or less than about 250 nm in diameter.In certain such embodiments, the nanoparticles of the present disclosuremay be between about 10 nm and about 200 nm, between about 30 nm andabout 100 nm, or between about 40 nm and about 80 nm in diameter. Asused herein.

Certain cytokines may cause or be involved in the process ofinflammation. For example the pro-angiogenic interleukin-1 (IL-1) may bepartially responsible for endogenous inflammatory cytokine release.Anti-angiogenic compounds such as tetrac and triac may be capable ofinhibiting pro-angiogenic various cytokines or chemokines. As a result,cytokines or chemokines-mediated angiogenesis contribute to thedevelopment of inflammation. The novel compound's inclusion ofanti-angiogenic thyroid hormone analogues such as tetrac, triac andnano-formulations thereof may assist in the suppression of cytokines andchemokines responsible for early stage inflammation which may precede anadaptive immune response by the body.

Interleukin responses to conjugated thyroid hormone analog, specificallyconjugated nanoparticulate tetrac (Nanotetrac™) were observed in humanbreast cancer (MDA-MB-231) cells. Referring to the results depicted inFIG. 1, exposure of MDA-MB-231 cells to the Nanotetrac™ actingexclusively at integrin αvβ3, demonstrated a reduction in IL-1α andIL-1β mRNA accumulation by 50-60%. Moreover, it was also determined thatthe Nanotetrac™ may also reduce interleukin-6 (IL-6) mRNA by 25%, andactually increased interleukin-11 (IL-11) mRNA abundance by 30%. Theseresults demonstrate a selective effect for reduction of thepro-inflammatory IL-1 and IL-6 while also increasing IL-11 which is notpro-inflammatory, but rather a desirable stimulator of hematopoieticstem cell proliferation.

Embodiments of the conjugated thyroid hormone analog, such asNanotetrac™ may also express selective anti-inflammatory effects towardchemokine receptors and chemokine ligand gene expression. For example,studies were performed measuring the effects of Nanotetrac™ on tumorcell expression of the mRNA of the chemokine ligand CX3CL1 (also knownas “fractalkine”) and the mRNA of its CX3CR1 receptor for fractalkine.Referring to FIG. 2, the results demonstrate that both the expression ofCX3CR1 receptor and its CX3CL1 ligand are concomitantly decreased byapproximately 75% in response to Nanotetrac™. Fractalkine may mediatechemotaxis and adhesion of inflammatory cells via its receptor.Furthermore, fractalkine may be considered a pharmacologically highpriority anti-inflammatory target, because fractalkine may participatein the early inflammatory components of several neurodegenerativediseases, including Alzheimer's disease, Parkinson's disease andHIV-associated encephalopathy.

Still referring to FIG. 2, the results of experimentation furtherindicate that the chemokine ligand CXCL10 and chemokine receptor CXCR4may be modestly increased by approximately 20% when cells are exposed tothe conjugated thyroid hormone analogue formulation. CXCL10 may beinvolved in the promotion of apoptosis and of angiostasis. The selectiveinduction of CXC10 may be desirable, and in certain settings thechemokine ligand and receptor can support cell growth and chemotaxis. Itis clear based on these findings, that the thyroid hormone receptor onαvβ3 may selectively mediate thyroid hormone analogue actions onchemokines and their receptors, and that the pharmaceutical targeting ofcertain chemokine systems may be feasible via αvβ3 and agents such asNanotetrac™.

The inhibition of the early inflammatory response (innate immuneresponse) with conjugated thyroid hormone analogue formulations such asNanotetrac™ may begin at αvβ3. There are certain actions of agonistthyroid hormones such as T4 and T3 that are relevant to the developmentand promotion of inflammation, which may be inhibited by conjugatedthyroid hormone analogues such as Nanotetrac™. For example, T4 and T3may act to modulate the production of Signal transducer and activator oftranscription-3 (STAT3) which transduces signals of a number ofinflammatory cytokines, such as interleukins, and the phosphorylation ofSTAT3. The regulation of STAT3 may occur by the introduction ofpro-angiogenic thyroid hormone, for example the introduction of apro-angiogenic thyroid hormone concentration such as T4 between 10⁻¹⁰ to10⁻⁷ M. The effect of T4 on STAT3 has been reproduced by T4 chemicallybonded to agarose (agarose-T4) (10⁻¹⁰ M free T4). The binding of T4 toagarose prevents the T4 from gaining access to the cell's interior. Themodulation of STAT3 while being unable to enter the cell's interiorindicates that the plasma membrane hormone receptor for thyroid hormoneon αvβ3 is involved. Conversely, STAT3's inflammatory response may bereduced or minimized by the introduction of an anti-angiogenic thyroidhormone.

Thyroid hormones may also potentiate certain effects of interferon-γ(IFN-γ) which may also induce certain chemokines as well as refinegrowth factor signaling at the epidermal growth factor (EGF) receptor(EGFR). It has been determined experimentally that there may becross-communication between EGFR, cytokine and chemokine signalingpathways that may be stimulated by agarose-T4. The stimulation of thesepathways may further implicate αvβ3 in the IFN-γ and EGFR, as well asSTAT3, behaviors described above and the promotion of the inflammatoryresponse. Nanotetrac™ and other conjugated thyroid hormone analogues mayinhibit the expression of the EGFR gene, thus reducing the crosscommunication between EGFR, cytokines and chemokine signal pathwaysultimately reducing the overall inflammatory response produced byendogenous thyroid hormone.

As used herein, the phrase “growth factors” or “neurogenesis factors”may refer to proteins, peptides or other molecules having a growth,proliferative, or trophic effect on cells of the CNS or PNS. Suchfactors may be used for inducing proliferation or differentiation andcan include, for example, any trophic factor that allows cells of theCNS or PNS to proliferate, including any molecule which binds to areceptor on the surface of the cell to exert a trophic orgrowth-inducing effect on the cell. Preferred factors include, but arenot limited to, nerve growth factor (“NGF”), epidermal growth factor(“EGF”), platelet-derived growth factor (“PDGF”), insulin-like growthfactor (“IGF”), acidic fibroblast growth factor (“aFGF” or “FGF-1”),basic fibroblast growth factor (“bFGF” or “FGF-2”), and transforminggrowth factor-alpha and -beta (“TGF-α” and “TGF-β”).

The integrin αvβ3 may be generously or overly expressed by tumor cellsand dividing blood vessel cells. As disclosed above, thyroid hormonessuch as tetrac and triac may bind exclusively to the αvβ3 integrinreceptor, making the thyroid hormone of the conjugated thyroid hormoneanalogue a selective targeting mechanism for tumor cells which expressthe integrin receptor. Furthermore, conjugated thyroid hormone analoguesor other ligands of αvβ3 may have significant potential either alone orin combination with other anti-inflammatory agents because integrin αvβ3is present on plasma membranes of the cells relevant to the formation ofinflammation. For example, integrin αvβ3 may be found on the plasmamembrane of neutrophils, peripheral blood lymphocytes, and alveolarmacrophages at the sites of lung inflammation.

The early inflammatory component of the innate immune response mayinclude contributions from inflammatory cells, response-modifyingcytokines and chemokines and blood vessel growth factors. With regard tothe latter, it was pointed out above that Nanotetrac™ may blockcontributions to the pro-angiogenic component of inflammation viaactions on interleukins as well as by decreasing the expression ofcytokine and chemokine mRNA. Vascular endothelial growth factor (VEGF),basic fibroblast growth factor (bFGF), platelet-derived growth factor(PDGF), insulin-like growth factor-1 (IGF-1) and EGF are factors thathave all been implicated in the vascular phase of the inflammatoryresponse. Acting via the receptor on integrin αvβ3, Nanotetrac™ andother conjugated thyroid hormone analogues may block the pro-angiogenicactions of each of these factors.

Iodothyronines may also modify activities of inflammatory cells whichhave been shown to express the thyroid hormone/tetrac receptor-bearingintegrin αvβ3. Acting via the cell surface receptor, thyroid hormone mayincrease reactive oxygen species (‘respiratory burst’) in granulocytes.Macrophage function may also be enhanced by thyroid hormone. The lattereffect has been thought to result from actions of thyroid hormone withinthe cell (genomic actions'), but it is clear that thyroid hormoneanalogues, such as Nanotetrac, act at αvβ3 to modulate function ofnuclear thyroid hormone receptors via control from the integrin offunctions of nuclear co-activator proteins and of phosphorylation ofnuclear receptor proteins.

Experimental data also suggests that the conjugated thyroid hormoneanalogues such as Nanotetrac™ formulations may further interfere withgene expression programs triggered in target cells by the increasedexpression of small non-coding trans-regulatory snpRNAs which areassociated with innate immunity/inflammation pathway activation in humancells. Activation of trans-regulatory non-coding snpRNA-associatedpathways has been linked with the engagement of the long-rangeintergenic enhancers and may be pathogenically associated with increasedrisk of developing prostate cancer and other common human disorders.

Furthermore, in some instances, there is crosstalk between thetetrac-thyroid hormone receptor on integrin αvβ3 and estrogen receptor-α(ERα) in human lung carcinoma cells that express this estrogen receptor.The proliferative effect of thyroid hormone at αvβ3 in such cells may bedependent upon ERα. This observation raises the possibility that theactions of thyroid hormone and anti-angiogenic agents on inflammationthat are mediated by their receptor on integrin αvβ3 may be involvedwith other non-peptide hormone response systems that may be regulated atthe cell surface.

In one or more embodiments, the conjugated thyroid hormone analogue mayfurther include one or more anti-inflammatory agents encapsulated by thepolymer. The anti-inflammatory encapsulated within the polymer may bereferred to as the “payload”. The amount of the payload the polymer mayencapsulate vary depending on the polymer being used and theanti-inflammatory agent being encapsulated. An anti-inflammatory agentmay be any substance that has a mechanism of action that reduces,partially reduces or suppresses inflammation. “Encapsulation” may referto one or more substances surrounding, encasing or protecting anothersubstance, from the environment. For example, in some embodiments, thepolymer may shield or protect an anti-inflammatory from harmfulconditions in the body that may prematurely break down or degrade theanti-inflammatory agent prior to reaching the target site. In someinstances, the anti-inflammatory agent may be encased or fullysurrounded by the polymer. In other embodiments, the anti-inflammatorymay be bound, attached, adsorbed, or bound to the polymer shielding itusing intermolecular forces such as dipole-dipole interactions,ion-dipole interaction, ion-induced interaction, hydrogen bonding,London-dispersion forces, Van der Waals forces, Keesom forces, or Debyeforces.

Polymeric microparticles and nanoparticles in some embodiments may beformulated by self-assembly of homopolymers or copolymers. The polymericparticle may include in some embodiments, alternating copolymers orblock copolymers consisting of two or more polymer chains with differinghydrophobicity. For instance, in some embodiments, the polymers orcopolymers may spontaneously assemble into a core-shell structure in anaqueous environment to minimize the system's free energy. In an instancewhere a block co-polymer is used, the hydrophobic blocks may form thecore to minimize exposure of the aqueous surroundings, whereas ahydrophilic block may form a shell to stabilize the core through directcontact with water.

There are several methods available for preparing polymer microparticlesand nanoparticles followed by encapsulating one or more agents, such asanti-inflammatory agents. Methods for preparing the polymeric particlesmay include emulsification-solvent preparation methods, including singleemulsion and double emulsion methods. Some embodiments may use methodssuch as nano-precipitation (also known as the solvent displacementmethod), the salting out method or by using microfluidic devices. Thechoice of the method for preparing the polymeric particle ornanoparticle for encapsulation may vary depending on the nature of thesubstance being encapsulated or entrapped within the polymeric particle.For the encapsulation of a hydrophilic substance, double emulsionmethods may be preferred, whereas for the encapsulation of a hydrophobicsubstance, nano-precipitation, single emulsion or salting out methodsmay be used. In some embodiments, the microfluidic device for theformation of polymeric particles may be implemented such as a situationwherein the conditions necessitate fast mixing in homogenous conditionson the micro scale. The release rates of drugs encapsulated inside canbe controlled by modifying the polymer's chemical and physicalproperties.

For example, in one embodiment, tetrac PLGA nanoparticles encapsulatingan NSAID and polyphenols were prepared using a double emulsion andsolvent evaporation method. A stock solution of PLGA-Tetracnanoparticles polymer was prepared by dispersing 100 mg/mL containing4-8% of Tetrac/PLGA (w/w) in dichloromethane. A stock solution of 10mg/mL of NSAID and/or polyphenol was prepared by dissolving the NSAIDsuch as ibuprofen and/or polyphenol such as resveratrol indichloromethane. Five hundred μL of each stock solution was mixedtogether by vortexing. Then, 1 mL of this solution was mixed with 200 μLof PBS by intermittent sonication (2-3 times, 30 sec each time) toobtain a primary emulsion. As depicted in FIG. 3, the primary emulsionwas then intermittently emulsified by sonication (30 s) in 2 mL of 1%w/v PVA solution. This water-in-oil-in-water emulsion was then added to40 mL of 1.0% PVA solution and stirred for 30 min under constantmagnetic stirring. Immediately after, dichloromethane was evaporated atlow pressure at 37° C. using a rotatory evaporator. The whole solutionwas then dialyzed using 10-12 KD dialysis membrane against water for 24hours to remove the impurities and residual solvents. The entiresolution was lyophilized and re-dispersed for further use.

The size distribution of the of the PLGA-tetrac nanoparticlesencapsulating resveratrol in an aqueous dispersion using the doubleemulsion and solvent evaporation method described above, was analyzed bydynamic light scattering (DLS) using a Malvern zeta sizer. After there-dispersion of the lyophilized powder in deionized water, 1 mL of theNP solution was taken in 3 mL of a four size clear plastic cuvette andmeasured directly by the DLS. Referring to the results depicted in FIG.4, the average size of PLGA-Tetrac NPs encapsulating resveratrol rangedfrom 150-250 nm.

The conjugated thyroid hormone analogue may deliver theanti-inflammatory agent locally to the site of inflammation as thethyroid hormone portion of the conjugated thyroid hormone analoguetargets the integrin receptor αvβ3. For example, a tetrac moietycovalently bound to a PLGA polymer may be used as a ligand of αvβ3,expressed by rapidly dividing endothelial cells at the sites ofinflammation. The anti-inflammatory agent may be encapsulated by thePLGA particle, thus as the tetrac selectively targets and seeks out theαvβ3 bearing endothelial cells, the PLGA nanoparticle may release theanti-inflammatory agent locally right at the point of inflammation.

The encapsulated anti-inflammatory agents within the polymer may beselected from non-steroidal anti-inflammatory drugs (NSAIDS),salicylates, anti-inflammatory glucocorticoids, anti-fibrotic agentsthat exhibit anti-inflammatory properties such as pirfenidone, CD-47antibodies or a combination of anti-inflammatory agents thereof. AnNSAID may be any group of anti-inflammatory and analgesic drugs that maysuppress inflammation and pain by inhibiting the cyclooxygenase pathwayand preventing release of inflammatory mediators (e.g. prostacyclin,prostaglandins and thromboxane). NSAIDs may bind to cyclooxygenase-1(COX-1), cyclooxygenase-2 (COX-2) or a combination of COX inhibitors. Insome embodiments, the NSAIDS encapsulated within the polymer of theconjugated thyroid hormone analogue may include but is not limited toibuprofen, diclofenac, and diclofenac with misprostol, indomethacin,ketoprofen, fenbrufen, naproxen, sulindac, celecoxib, nabumetone,mefenamic acid, oxyphenbutazone, diflunisal, etodolac, fenoprofen,flurbiprofen, meclofenamate, meloxicam, nabumetone, oxaprozin,piroxicam, tolmetin, valdecoxib and propionic acid derivatives.

In another embodiment, the anti-inflammatory agent may include one ormore salicylates encapsulated by the polymer of the conjugated thyroidhormone analogue. A salicylate may be a salt or ester of salicylic acid(C₆H₄(OH)COOH). Salicylates may have an OH group in the ortho positionto the carboxyl group. In some instances, a salicylate may be referredto as 2-hydroxybenzoic acid. Similar to the NSAIDs, the salicylates mayreduce the onset and extent of inflammation by inhibiting thecyclooxygenase enzyme (COX) production, including both COX-1 and COX-2.Salicylates may include one or more of the following compoundsencapsulated inside the polymer of the conjugated thyroid hormoneanalog, including but not limited to aspirin, choline salicylate,choline and magnesium salicylate, magnesium salicylate, and sodiumsalicylate.

Embodiments of the conjugated thyroid hormone analogue comprising one ormore anti-inflammatory glucocorticoids may include one or more of thefollowing compounds encapsulated by the polymer of the conjugatedthyroid hormone analogue. A glucocorticoid may be any corticoidsubstance that increases gluconeogenesis and may raise the concentrationof glycogen in the liver and blood glucose. An anti-inflammatoryglucocorticoid may be any glucocorticoid that has an effect on theinflammation response by the body, for example by inhibiting the releaseof histamine. The glucocorticoids may bind to glucocorticoid receptorsin the cytoplasm which then dimerize and translocate to the nucleus,where they bind to glucocorticoid response elements (GRE) onglucocorticoid-responsive genes, resulting in increased transcription.Glucocorticoids may increase the transcription of genes coding foranti-inflammatory proteins, including lipocortin-1, interleukin-10,interleukin-1 receptor antagonist and neutral endopeptidases. Theanti-inflammatory response may be due to a direct inhibitory interactionbetween activated glucocorticoid receptors and activated transcriptionfactors, such as nuclear factor-kappa B and activator protein-1, whichregulate the inflammatory gene expression. The glucocorticoids may alsoinhibit the expression of multiple inflammatory genes such as cytokines,enzymes, receptors and adhesion molecules. Glucocorticoid receptors mayalso interact with CREB-binding protein (CBP), which may act as aco-activator of transcription, binding several other transcriptionfactors that compete for binding sites on this molecule. Increasedtranscription may be associated with uncoiling of DNA wound aroundhistone and this may be secondary to acetylation of the histone residuesby the enzymatic action of CBP. Glucocorticoids may lead todeacetylation of histone, resulting in tighter coiling of DNA andreduced access of transcription factors to their binding sites, therebysuppressing gene expressions that may lead to inflammation. Saidanti-inflammatory glucocorticoids may include, but are not limited tohydrocortisone, cortisone, cortisol, dexamethasone, dexamethasoneIntensol™, budesonide, methylprednisolone, prednisolone, prednisolonesodium phosphate and prednisone.

In some embodiments, the anti-inflammatory agent being encapsulated bythe polymer may include anti-fibrotic agents having anti-inflammatoryproperties. An anti-fibrotic agent may be an agent that causes theregression of fibrosis. An example of an anti-fibrotic agent withanti-inflammatory effects may include pirfenidone, NOS-2, daidzein,sirolimus and tyrosine kinase inhibitors including nintendanib.

Embodiments of the anti-inflammatory agent being encapsulated by thepolymer may include anti CD47 antibodies. An anti-CD47 antibody may bean anti-body such as a monoclonal or polyclonal antibody that binds toan integrin associated transmembrane protein encoded by the CD47 gene.By inhibiting the CD47 gene with an anti-CD47 antibody, may reduceinflammation by reducing the recruitment of T-cells by the body as wellas neutrophils and monocytes at the area of inflammation. Examples ofanti-CD47 antibodies may include B6H12.2 (ab3283), EPR4150 (ab108415),ab175388, OX-101 (33852), MEM-122 (ab9089), ab136550, Allophycocyanin(ab134485), ab118222, ab171767, ab176099, ab174029 and human CD47protein fragment ab151372.

In some embodiments, the polymer of the conjugated thyroid hormoneanalogue may further include encapsulated anti-oxidant polyphenolsinside the polymer for local release at the site of inflammation. Apolyphenol may refer to a compound containing one or more phenolichydroxyl groups. An anti-oxidant polyphenol may be a polyphenol thatprevents or inhibits oxidation or reactions promoted by oxidants, suchas oxygen, peroxide or free radicals. The anti-oxidant polyphenol mayinclude one or more flavones, isoflavones and/or flavonoids such asresveratrol, quercetin, myricetin, catechin, epigallocatechin, enisteinand combinations thereof.

In yet another embodiment, the polymer may encapsulate one or moreadditional agents to release at thyroid hormone analogue target bindingsite integrin αvβ3. One or more additional agents that may beencapsulated within the polymer may include, but is not limited to,bisphosphonates such as risendronate, alendronate, ibandronate,etidronate, pamidronate, tiludronate, and zoledronic acid, growthfactors, hormones, enzymes, antibiotics, vasodilators, anti-coagulants,anti-virals, anti-bacterials, immuno-suppressants, analgesics,vascularizing agents, or cell adhesion molecules, or combinationsthereof or other bioactive agents.

In some embodiments, the conjugated thyroid hormone analogue, includingone or more additionally encapsulated agents may be administeredsystemically to one or more areas of inflammation or reactiveangiogenesis, wherein the integrin receptor may be expressed. Theconjugated thyroid hormone analogue, including one or more encapsulatedagents, may be administered at a therapeutic concentration ofapproximately 200-2000 μg/day. In another embodiment, the concentrationmay vary between 200-1800 μg/day, 300-1700 μg/day, 500-1500 μg/day,700-1200 μg/day or 800-1000 μg/day.

Nanoparticles within the present disclosure may include up toapproximately 100, up to 90, up to 80, up to 70, up to 60, up to 50, upto 40 up to 30, up to 20 or up to 10 molecules of thyroid hormoneanalogues conjugated per nanoparticle. By way of non-limiting example,the ratio of the thyroid hormone analogues or other therapeuticmolecules per nanoparticle may range from a ratio of 1 thyroid hormoneanalogue molecule per 1 nanoparticle (shown also as 1:1) up to 100thyroid hormone analogues per nanoparticle (shown also as 100:1). Morepreferably, the range may be from 10:1-50:1 (i.e., 10:1, 20:1, 30:1,40:1) thyroid hormones or thyroid hormone analogues or other therapeuticmolecules per nanoparticle. In other embodiments, the ratio ofconjugated thyroid hormone analogues may range from 10:1-50:1,30:1-40:1, 20:1-25:1 or 10:1-20:1 thyroid hormone analogue molecules pernanoparticle. In various embodiments, the density of the thyroid hormoneanalogues in the nanoparticles is between 0.1 and 25%, for example thedensity of the thyroid hormone analogue particle may be betweenapproximately 0.1-1%, 0.5-2%, 1-3%, 2-5%, 3-7%, 4-10%, 5-15%, 7-20%,0.2-25%, 0.5-20%, 1-20% or 1-15%.

In some embodiments, the nanoparticles within the present disclosure mayinclude up to approximately 100, up to 90, up to 80, up to 70, up to 60,up to 50, up to 40 up to 30, up to 20 or up to 10 molecules ofanti-inflammatory agents per nanoparticle. By way of non-limitingexample, the ratio of the anti-inflammatory or other agents pernanoparticle may range from a ratio of 1 anti-inflammatory molecule per1 nanoparticle (shown also as 1:1) up to 100 anti-inflammatory agentmolecules per nanoparticle (shown also as 100:1). More preferably, therange may be from 10:1-30:1 (i.e., 10:1-30:1) anti-inflammatory agentsor other therapeutic molecules per nanoparticle. In other embodiments,the ratio of anti-inflammatory agents may range from 10:1-50:1,30:1-40:1, 20:1-25:1 or 10:1-20:1 anti-inflammatory agents pernanoparticle. In various embodiments, the density of theanti-inflammatory agent in the nanoparticles may be between 0.1 and 25%,for example the density of the anti-inflammatory agent may be betweenapproximately 0.1-1%, 0.5-2%, 1-3%, 2-5%, 3-7%, 4-10%, 5-15%, 7-20%,0.2-25%, 0.5-20%, 1-20% or 1-15%. In some embodiments, theanti-inflammatory agent may include at least 10 molecules ofanti-inflammatory agent per nanoparticle or microparticle polymer. Forexample, the polymeric particle may encapsulate at least 10, at least20, at least 30, at least 40, at least 50, at least 100, at least 200,at least 500 anti-inflammatory molecules per nanoparticle.

In some embodiments, the anti-inflammatory NSAIDs may be administered atany known therapeutic concentration. For example, therapeuticconcentrations may be approximately 50 μg-2000 mg/day. The concentrationand effective dose may vary depending on the NSAID being encapsulated.For example, the concentration may vary between 50-200 μg/day, 200-500μg/day, 500-1000 μg/day, 1-50 mg/day, 50-100 mg/day, 100-200 mg/day,200-400 mg/day, 400-800 mg/day, 800-1000 mg/day or 1000-2000 mg/day. Insome embodiments, the NSAID dose or payload may be administered between1-1000 mg, 1-500 mg, 1-100 mg, 1-50 mg or 1-10 mg. In other embodiments,the dose may be significantly lower and may range from 50-100 μg, 50-500μg, or 50-1000 μg.

Embodiments, of the thyroid hormone analogues that may includesalicylates encapsulated by the polymeric particles, may be administeredat any known therapeutic concentration. For example, salicylates may beadministered at a therapeutic concentration between approximately 50μg-6000 mg/day. The concentration and effective dose may vary dependingon the salicylate being encapsulated and administered. For example, theconcentration may vary between 50-200 μg/day, 200-500 μg/day, 500-1000μg/day, 1-50 mg/day, 50-100 μg/day, 100-200 mg/day, 200-400 mg/day,400-800 mg/day, 800-1000 mg/day, 1000-2000 mg/day, 2000-3500 mg/day,3500-5000 mg/day or 5000-6000 mg/day. In some embodiments, the dose orpayload of salicylates may be administered between 1-1000 mg, 1-500 mg,1-100 mg, 1-50 mg or 1-10 mg. In other embodiments, the dose may besignificantly lower and range from 50-100 μg, 50-500 μg, or 50-1000 μg.

Embodiments, of the thyroid hormone analogues that may includeanti-inflammatory glucocorticoids encapsulated by the polymericparticles, may be administered at any known therapeutic concentration.For example, anti-inflammatory glucocorticoids may be administered at atherapeutic concentration between approximately 1 μg-100 mg/day. Theconcentration and effective dose may vary depending on whichanti-inflammatory glucocorticoids is being encapsulated andadministered. For example, the concentration may vary between 1-50μg/day, 50-100 μg/day, 100-500 μg/day, 500-1000 μg/day, 1-5 mg/day, 5-10mg/day, 10-20 mg/day, 20-25 mg/day, or 25-30 mg/day, 30-100 mg/day. Insome embodiments, the dose or payload of anti-inflammatoryglucocorticoids may be administered between 1-100 mg, 1-50 mg, or 1-10mg. In other embodiments, the dose may be significantly lower and rangefrom 50-100 μg, 50-500 μg, or 50-1000 μg.

In some embodiments that include anti-oxidant polyphenols encapsulatedby the polymeric particles, the anti-oxidant polyphenols may beadministered at any known therapeutic concentration. The therapeuticconcentration of the encapsulated anti-oxidant polyphenols may vary from1 μg-5000 mg/day depending on the anti-oxidant polyphenol beingadministered. For example, the anti-oxidant polyphenol may beadministered in therapeutic concentration between approximately 1-50μg/day, 50-100 μg/day, 100-500 μg/day, 500-1000 μg/day, 1-100 mg/day,100-300 mg/day, 300-600 mg/day, 600-800 mg/day, 800-1000 mg/day,1000-2000 mg/day, 2000-3500 mg/day or 3500-5000 mg/day. In someembodiments, the dose or payload of anti-oxidant polyphenols may beadministered between 1-5000 mg, 1-3000 mg, 1-1000 mg, 1-500 mg, 1-100mg, 1-50 mg or 1-10 mg. In other embodiments, the dose may besignificantly lower and range from 1-50 μg, 50-100 μg, 50-500 μg, or50-1000 μg.

Embodiments including anti-fibrotic agents having anti-inflammatoryproperties encapsulated by the conjugated thyroid hormone analogue maybe administered at a dose between 1 mg-3500 mg/day. For example, theanti-fibrotic agent may be administered in therapeutic concentrationbetween approximately 1-50 mg/day, 50-100 mg/day, 100-500 mg/day,500-1000 mg/day, 1000-2000 mg/day or 2000-3500 mg/day. In someembodiments, the dose or payload of anti-fibrotic agents havinganti-inflammatory properties may be administered between 1-3500 mg,1-3000 mg, 1-1000 mg, 1-500 mg, 1-100 mg, 1-50 mg or 1-10 mg. In otherembodiments, the dose may be significantly lower and range from 1-50 μg,50-100 μg, 50-500 μg, or 50-1000 μg.

Embodiments including anti-CD47 antibodies encapsulated by theconjugated thyroid hormone analogue may be administered at a dosebetween 1 μg/day-500 mg/day. For example, the anti-CD47 antibody may beadministered in therapeutic concentration between approximately 1-50μg/day, 50-100 μg/day, 100-500 μg/day, 500-1000 μg/day, 1-200 mg/day or200-500 mg/day. In some embodiments, the dose or payload of theanti-CD47 antibody may be administered between 1 μg-500 mg, 1-30 μg,1-100 μmg, 1-500 μg, 500-1000 μg, 1-100 mg, 1-50 mg or 1-10 mg.

The use of the encapsulation into the conjugated thyroid hormoneanalogue and precise targeting of the anti-inflammatory agent to theinflammation site produces unexpected results in the therapeutic doseadministered to control, reduce or suppress the inflammation. Theencapsulation of the anti-inflammatory agent in the polymer may allowfor lower doses of the anti-inflammatory agent to be administered thanin a situation wherein the anti-inflammatory were administered on itsown. In particular, doses administered between 1 μg-100 mg or 1 mg-100mg of the encapsulated anti-inflammatory may be significantly less thanthe therapeutic dose administered without being encapsulated. Forexample, a common NSAID such as Ibuprofen may have a therapeutic dosethat is 200-600 mg when administered without targeted administration inthe encapsulated polymer. When encapsulated, the ibuprofen may reduceinflammation in extremely low doses between 1-100 mg, which is between16-50% of the therapeutic dose when 100 mg is administered and 0.2-0.5%of the typical therapeutic dose when 1 mg is administered. Ultimately,lowered doses may mean less toxicity and adverse side effects. Forinstance, in the case of ibuprofen, decreased instances of ulcers,bleeding, headaches, nausea, diarrhea, abdominal pain etc.

The conjugated thyroid hormone analogue may be administered with one ormore pharmaceutically acceptable carriers. “Pharmaceutically acceptablecarriers” may refer to and include any and all solvents, dispersionmedia, coatings, antibacterial and antifungal agents, isotonic andabsorption delaying agents, and the like which are compatible with theactivity of the compound and are physiologically acceptable to thesubject. An example of a pharmaceutically acceptable carrier is bufferednormal saline (0.15M NaCl). The use of such media and agents forpharmaceutically active substances is well known in the art. Exceptinsofar as any conventional media or agent is incompatible with thetherapeutic compound, use thereof in the compositions suitable forpharmaceutical administration is contemplated. Supplementary activecompounds can also be incorporated into the compositions.

In some embodiments, the conjugated thyroid hormone analogue,encapsulating one or more additional agents may be administered directlyinto the site of inflammation. For example, the composition may beinjected into inflamed joints and muscles. In other embodiments, theconjugated thyroid hormone analogue may be formulated for administrationvia one or more of the following routes, including but not limited toparenteral including via catheterization, intravenous, oral, rectal,topical such as a Band-Aid, cream, ointment or a gauze pad, ophthalmic,local implantation, subcutaneous, intramolecular, intraperitoneal,intramuscular, buccal, vaginal, intraorbital, intracerebral,intracranial, intraspinal, intraventricular, intrathecal,intracisternal, intracapsular, intranasal or by aerosol administrationor a combination of routes thereof.

For enteral administration, a compound can be incorporated into an inertcarrier in discrete units such as capsules, cachets, tablets orlozenges, each containing a predetermined amount of the active compound;as a powder or granules; or a suspension or solution in an aqueousliquid or non-aqueous liquid, e.g., a syrup, an elixir, an emulsion or adraught. Suitable carriers may be starches or sugars and includelubricants, flavorings, binders, and other materials of the same nature.

In some embodiments, a medical device may be coated with the conjugatedthyroid hormone analogue wherein the polymer has encapsulated at leastone additional anti-inflammatory agent. The coated medical device mayinclude stents, catheters, cannulas or electrodes.

In another embodiment, conjugated thyroid hormone analogues may furtherencapsulate one or more nerve growth factors or other neurogenesisfactors useful to protect against damage associated with the body'simmune/inflammatory response to an initial injury to nerve tissue. Sucha response may follow trauma to nerve tissue, caused, for example, by anautoimmune dysfunction, neoplastic lesion, infection, chemical ormechanical trauma, disease, by interruption of blood flow to the neuronsor glial cells, or by other trauma to the nerve or surrounding material.For example, the primary damage results from hypoxia orischemia-reperfusion following occlusion of a neural blood supply, as inan embolic stroke, is believed to be immunologically associated. Inaddition, at least part of the damage associated with a number ofprimary brain tumors also appears to be immunologically related.Application of a polymeric thyroid hormone analogue alone or incombination with nerve growth factors or other neurogenesis factors,either directly or systemically alleviates and/or inhibits theimmunologically related response to a neural injury.

Alternatively, administration of an agent capable of stimulating theexpression and/or secretion in vivo of conjugated thyroid hormoneanalogues alone or in combination with nerve growth factors or otherneurogenesis factors expression, preferably at the site of injury, mayalso be used. Where the injury is to be induced, such as during surgeryor other aggressive clinical treatment, the conjugated thyroid hormoneanalogues alone or in combination with nerve growth factors or otherneurogenesis factors or agent may be provided prior to induction of theinjury to provide a neuroprotective effect to the nerve tissue at risk.

In some embodiments, the conjugated thyroid hormone analogue, includingone or more additionally encapsulated agents may be encapsulated into ahydrogel for local implantation into the inflammation site. Synthetichydrogels from methacrylate derived polymers may be used in biomedicalapplications because of their similarity to the living tissues. The mostwidely used synthetic hydrogels are polymers of acrylic acid, acrylamideand 2-hydroxyethyl methacrylate (HEMA). The poly HEMA may beinexpensive, biocompatible, available primary alcohol side chainelongation functionality for conjugation and fit for ocular, intraocularand other ophthalmic applications which make them perfect drug deliverymaterials. The poly HEMA may be immune to cell attachment and provideszero cell motility which makes them an ideal candidate for internaldelivery system. In one example, formulations of a hydrogel may includebiodegradable polymeric hydrogels, such as those disclosed in U.S. Pat.No. 5,410,016 to Hubbell et al. These polymeric hydrogels can bedelivered to the inside of a tissue lumen and the active compoundsreleased over time as the polymer degrades.

In addition to the aforementioned ingredients, formulations of theconjugated thyroid hormone analogue may further include one or moreoptional accessory ingredient(s) utilized in the art of pharmaceuticalformulations, e.g., diluents, buffers, flavoring agents, binders,surface active agents, thickeners, lubricants, suspending agents,preservatives (including antioxidants), excipients, dispersing agents;inert diluents, granulating and disintegrating agents, sweeteningagents, coloring agents, physiologically degradable compositions such asgelatin; aqueous vehicles and solvents; oily vehicles and solvents;dispersing or wetting agents; emulsifying agents, demulcents, buffers,salts, fillers, emulsifying agents, antioxidants, antibiotics,antifungal agents, stabilizing agents, and pharmaceutically acceptablepolymeric or hydrophobic materials and the like.

In an alternative embodiment, the release of a conjugated thyroidhormone analogue encapsulating one or more additional agent inside thepolymer, may be controlled by further encapsulating the polymer itselfwithin a liposome, microparticle, or nanoparticle. The breakdown of theliposome, microparticle or nanoparticle may be calculated to furthercontrol length of time wherein the conjugated thyroid hormone analogueis released at the site of inflammation. This embodiment may includeprolonged release times of the conjugated thyroid hormone betweenapproximately 1-72 hours, 10-60 hours, 15-50 hours, 20 to 40 hours orfor 24 hours.

Embodiments of the conjugated thyroid hormone analogue may further beapplied toward embodiments of methods for treating inflammatoryconditions. Inflammatory conditions that may be treated may include butis not limited to Parkinson's disease, trauma, cerebral ischemia,amyotrophic lateral sclerosis, multiple sclerosis, arthritis, myositis,poikiloderma, rosacea, psoriasis, acne, pityriasis rosea, eczema, and acombination of inflammatory conditions thereof. Furthermore, embodimentsof the method for treating an inflammatory condition may further includetreating musculoskeletal conditions that may or may not includeinflammation. The musculoskeletal condition may include conditions suchas aches and pains that are in the body's muscles, joints, tendons,ligaments, nerves, and combination of musculoskeletal conditionsthereof.

In one embodiment, the steps for treating an inflammatory condition mayinclude conjugating an anti-angiogenic thyroid hormone analogue such astetrac or triac to a polymer where the results of the conjugation mayform a conjugated thyroid hormone analog. Embodiments of methods fortreating an inflammatory condition may further comprise encapsulatinginside the polymer of the conjugated thyroid hormone analog, at leastone of the following, including but not limited to at least one NSAID,at least one salicylate, at least one anti-inflammatory glucocorticoid,an anti-fibrotic agent having anti-inflammatory properties such aspirfenidone or a combination of anti-inflammatory agents. Embodiments ofthe method may also include binding the conjugated thyroid hormoneanalog, with or without the presence of an additional agent encapsulatedwithin the polymer, to one or more of the following inflammationmodulating receptors such as cytokine receptors, interleukin receptorschemokine receptors or a combination of receptors thereof.

Embodiments of the method for treating an inflammatory condition myfurther comprise the additional step of encapsulating the polymer insidea liposome, microparticle or nanoparticle for example in an effort tocontrol the release of the conjugated thyroid hormone analogue. In someembodiments, the liposome or microparticle can be lodged in capillarybeds surrounding ischemic tissue, or applied to the inside of a bloodvessel via a catheter.

Moreover, the method for treating one or more inflammatory conditionsmay also include the additional step of administering the conjugatedthyroid hormone analog, with or without the additional encapsulatedanti-inflammatory agent, at a therapeutic concentration betweenapproximately 200 μg/day to approximately 1000 mg. In some embodimentsthe dose administered may be less than 1000 mg/day, less than 500mg/day, less than 200 mg/day, less than 50 mg/day, less than 2000μg/day, less than 1500 μg/day, less than 1000 μg/day, less than 500μg/day or less than 200 μg/day. The dosage administered may varydepending on dosing factors known to those skilled in the art. Theperiod of dosing may vary depending on the dosing amount beingadministered. The step of administering may be performed at a specificinterval of time, for example, the step of administration may occur atan interval of once a week, once per day (i.e. every 24 hours), onceevery other day (i.e. every 48 hours), once every three days (i.e. every72 hours) or twice a day or more.

The step of administering the conjugated thyroid hormone, with orwithout the additional encapsulated anti-inflammatory agent may occurtopically, parenterally, locally at the site of inflammation,systemically by injectable routes such as subcutaneous, intravenous,intraperitoneal, intramuscular, intracerebral, intraorbital,intracranial, intraspinal, intraventricular, intrathecal,intracisternal, intracapsular, or by catheterization, orally, rectally,ophthalmically, local implantation, buccal, vaginal, intranasal or byaerosol administration or a combination of routes thereof.

While this disclosure has been described in conjunction with thespecific embodiments outlined above, it is evident that manyalternatives, modifications and variations will be apparent to thoseskilled in the art. Accordingly, the embodiments of the disclosure asset forth above are intended to be illustrative, not limiting. Variouschanges may be made without departing from the spirit and scope of theinvention as defined in the following claims. The claims provide thescope of the coverage of the invention and should not be limited to thespecific examples provided herein.

What is claimed:
 1. A composition comprising: a thyroid hormone analogueconjugated to a polymer, wherein the thyroid hormone analogue isimmobilized to a surface of the polymer, wherein said thyroid hormoneanalogue targets integrin αvβ3; and at least one anti-inflammatory agentencapsulated by the polymer, wherein said at least one anti-inflammatoryagent is selected from a non-steroidal anti-inflammatory drug (NSAID), atyrosine kinase inhibitor, a salicylate, an anti-inflammatoryglucocorticoid and an anti-fibrotic agent having anti-inflammatoryproperties.
 2. The composition of claim 1, wherein the thyroid hormoneanalogue is at least one of tetraiodothyroacetic acid (tetrac) andtriiodothyroacetic acid (triac).
 3. The composition of claim 1, whereinthe polymer is selected from the group consisting of polylactic acid(PLLA), polyglycolic acid (PGA), polyacrylic acid,poly(lactic-co-glycolic acid) (PLGA), polyethylene glycol (PEG),poly-L-lysine, chitosan, hyaluronic acid, fatty acids, polyamine,co-polymers thereof and combinations thereof.
 4. The composition ofclaim 1, wherein the polymer is formulated into a nanoparticle.
 5. Thecomposition of claim 1, wherein the NSAID is selected from the groupconsisting of ibuprofen, diclofenac, indomethacin, ketoprofen, naproxen,sulindac, celecoxib, nabumetone, mefenamic acid, oxyphenbutazone, andcombinations thereof.
 6. The composition of claim 1, further comprisingan anti-oxidant polyphenol encapsulated within the polymer.
 7. Thecomposition of claim 6, wherein the anti-oxidant polyphenol is at leastone of a flavone, an isoflavone, and a flavonoid.
 8. The composition ofclaim 7, wherein the flavonoid is selected from the group consisting ofresveratrol, quercetin, myricetin, catechin, epigallocatechin, genisteinand combinations thereof.
 9. The composition of claim 4, wherein thenanoparticle has a diameter between approximately 10 nm and <1000 nm.10. The method of claim 1, wherein the anti-fibrotic agents havinganti-inflammatory properties is pirfenidone.
 11. The composition ofclaim 1, further comprising one or more bisphosphonates encapsulated inthe polymer.
 12. The composition of claim 11, wherein the one or morebisphosphonates are selected from the group consisting of risendronate,alendronate, ibandronate, etidronate, pamidronate, tiludronate, andzoledronic acid.
 13. The composition of claim 1, wherein the compositionis formulated into a hydrogel.
 14. A method for making the compositionof claim 1 comprising the steps of: conjugating a thyroid hormoneanalogue to a polymer, forming a conjugated thyroid hormone analog; andencapsulating inside the polymer of the conjugated thyroid hormoneanalogue at least one of a non-steroidal anti-inflammatory drug (NSAID),a salicylate, an anti-inflammatory glucocorticoid and an anti-fibroticagent having anti-inflammatory properties.
 15. The method of claim 14,wherein the step of encapsulating includes encapsulating the at leastone of the non-steroidal anti-inflammatory drug (NSAID), the salicylate,the anti-inflammatory glucocorticoid and the anti-fibrotic agent havinganti-inflammatory properties at a dose between 1.0-100 mg.
 16. Themethod of claim 14, further comprising the step of encapsulating thepolymer in a liposome, microparticle or nanoparticle.
 17. The method ofclaim 14, wherein the NSAID is selected from the group consisting ofibuprofen, diclofenac, indomethacin, ketoprofen, naproxen, sulindac,celecoxib, nabumetone, mefenamic acid, oxyphenbutazone, and combinationsthereof.
 18. The method of claim 14, further comprising the step ofencapsulating at least one anti-oxidant polyphenol inside the polymer,wherein the anti-oxidant polyphenol is at least one of a flavone, anisoflavone, and a flavonoid.
 19. The method of claim 18, wherein theflavonoid is selected from the group consisting of resveratrol,quercetin, myricetin, catechin, epigallocatechin, enistein andcombinations thereof.
 20. The method of claim 14, wherein the polymer isselected from the group consisting of polylactic acid (PLLA),polyglycolic acid (PGA), polyacrylic acid, polyethylene glycol (PEG),poly-L-lysine, chitosan, hyaluronic acid, fatty acids, polyamine,co-polymers thereof and combinations thereof.
 21. A method comprisingadministering the composition of claim 1 to a subject.
 22. The method ofclaim 21, wherein the step of administering is performed at an intervalselected from the group consisting of once a day, every other day andonce a week.
 23. The method of claim 21, wherein the step ofadministering occurs topically, systemically, orally, locally at thesite of inflammation, and a combination thereof.